ABSTRACT
Systemic lupus erythematosus (SLE) is a complex systemic autoimmune disease driven by both innate and adaptive immune cells. African Americans tend to present with more severe disease at an earlier age compared with patients of European ancestry. In order to better understand the immunological differences between African American and European American patients, we analyzed the frequencies of B cell subsets and the expression of B cell activation markers from a total of 68 SLE patients and 69 normal healthy volunteers. We found that B cells expressing the activation markers CD86, CD80, PD1, and CD40L, as well as CD19+CD27-IgD- double-negative B cells, were enriched in African American patients vs. patients of European ancestry. In addition to increased expression of CD40L, surface levels of CD40 on B cells were lower, suggesting the engagement of the CD40 pathway. In vitro experiments confirmed that CD40L expressed by B cells could lead to CD40 activation and internalization on adjacent B cells. To conclude, these results indicate that, compared with European American patients, African American SLE patients present with a particularly active B cell component, possibly via the activation of the CD40/CD40L pathway. These data may help guide the development of novel therapies.
Subject(s)
B-Lymphocytes/cytology , Lupus Erythematosus, Systemic/ethnology , Black or African American , Antigens, Surface/analysis , B7-2 Antigen/analysis , CD40 Antigens/analysis , CD40 Ligand/analysis , Humans , PhenotypeABSTRACT
An empirical approach to improve the microsomal stability and CYP inhibition profile of lead compounds 1a and 1b led to the identification of 5 (BMS-341) as a dissociated glucocorticoid receptor modulator. Compound 5 showed significant improvements in pharmacokinetic properties and, unlike compounds 1a-b, displayed a linear, dose-dependent pharmacokinetic profile in rats. When tested in a chronic model of adjuvant-induced arthritis in rat, the ED50 of 5 (0.9 mg/kg) was superior to that of both 1a and 1b (8 and 17 mg/kg, respectively).
Subject(s)
Arthritis, Experimental/drug therapy , Cytochrome P-450 Enzyme Inhibitors/pharmacology , Heterocyclic Compounds, 3-Ring/pharmacology , Receptors, Glucocorticoid/metabolism , Thiadiazoles/pharmacology , Animals , Blood/drug effects , Blood/metabolism , Chemistry Techniques, Synthetic , Cytochrome P-450 CYP3A , Cytochrome P-450 CYP3A Inhibitors/chemistry , Cytochrome P-450 CYP3A Inhibitors/pharmacology , Cytochrome P-450 Enzyme Inhibitors/chemistry , Cytochrome P-450 Enzyme Inhibitors/pharmacokinetics , Disease Models, Animal , Dogs , Dose-Response Relationship, Drug , Drug Evaluation, Preclinical/methods , Drug Stability , Heterocyclic Compounds, 3-Ring/chemistry , Heterocyclic Compounds, 3-Ring/pharmacokinetics , Humans , Male , Rats, Inbred Lew , Receptors, Glucocorticoid/agonists , Structure-Activity Relationship , Thiadiazoles/chemistry , Thiadiazoles/pharmacokinetics , Transcription Factor AP-1/metabolismABSTRACT
Acylureas and acyclic imides are found to be excellent isosteres for 2-acylamino-1,3,4-thiadiazole in the azaxanthene-based series of glucocorticoid receptor (GR) agonists. The results reported herein show that primary acylureas maintain high affinity and selectivity for GR while providing improved CYP450 inhibition and pharmacokinetic profile over 2-acylamino-1,3,4-thiadiazoles. General methods for synthesis of a variety of acylureas and acyclic imides from a carboxylic acid were utilized and are described.
Subject(s)
Drug Discovery , Heterocyclic Compounds, 3-Ring/pharmacology , Receptors, Glucocorticoid/agonists , Thiadiazoles/pharmacology , Urea/pharmacology , Crystallography, X-Ray , Dose-Response Relationship, Drug , Heterocyclic Compounds, 3-Ring/chemical synthesis , Heterocyclic Compounds, 3-Ring/chemistry , Humans , Models, Molecular , Molecular Structure , Stereoisomerism , Structure-Activity Relationship , Thiadiazoles/chemistry , Urea/analogs & derivatives , Urea/chemistryABSTRACT
A series of heterocyclic glucocorticoid receptor (GR) modulators with 2,2-dimethyl-3-phenyl-N-(thiazol or thiadiazol-2-yl)propanamide core are described. Structure-activity relationships suggest a combination of H-bond acceptor and a 4-fluorophenyl moiety as being important structural components contributing to the glucocorticoid receptor binding and functional activity for this series of GR modulators.
Subject(s)
Amides/chemistry , Heterocyclic Compounds/chemistry , Receptors, Glucocorticoid/agonists , Thiadiazoles/chemistry , Thiazoles/chemistry , Heterocyclic Compounds/chemical synthesis , Heterocyclic Compounds/metabolism , Protein Binding , Receptors, Glucocorticoid/metabolism , Stereoisomerism , Structure-Activity RelationshipABSTRACT
Modification of a phenolic lead structure based on lessons learned from increasing the potency of steroidal glucocorticoid agonists lead to the discovery of exceptionally potent, nonsteroidal, indazole GR agonists. SAR was developed to achieve good selectivity against other nuclear hormone receptors with the ultimate goal of achieving a dissociated GR agonist as measured by human in vitro assays. The specific interactions by which this class of compounds inhibits GR was elucidated by solving an X-ray co-crystal structure.
Subject(s)
Amides/chemistry , Amides/pharmacology , Drug Discovery , Receptors, Glucocorticoid/agonists , Binding Sites , Crystallography, X-Ray , Humans , Indazoles/chemistry , Indazoles/pharmacology , Molecular Structure , Protein Binding/drug effects , Steroids/chemistry , Steroids/pharmacology , Structure-Activity RelationshipABSTRACT
SAR was used to further develop an indazole class of non-steroidal glucocorticoid receptor agonists aided by a GR LBD (ligand-binding domain)-agonist co-crystal structure described in the accompanying paper. Progress towards discovering a dissociated GR agonist guided by human in vitro assays biased the optimization of this compound series towards partial agonists that possessed excellent selectivity against other nuclear hormone receptors.
Subject(s)
Indazoles/chemical synthesis , Indazoles/pharmacology , Receptors, Glucocorticoid/agonists , Amides/chemistry , Amides/pharmacology , Humans , Indazoles/chemistry , Models, Molecular , Protein Binding/drug effects , Receptors, Glucocorticoid/chemistry , Structure-Activity Relationship , Sulfonamides/chemistry , Sulfonamides/pharmacology , Urea/chemistry , Urea/pharmacologyABSTRACT
Structurally novel 5H-chromeno[2,3-b]pyridine (azaxanthene) selective glucocorticoid receptor (GR) modulators have been identified. A screening paradigm utilizing cellular assays of GR-mediated transrepression of proinflammatory transcription factors and transactivation of GR-dependent genes combined with three physiologically relevant assays of cytokine induction in human whole blood has allowed for the identification of high affinity, selective GR ligands that display a broad range of pharmacological profiles. Agonist efficacy in reporter assays can be tuned by halogenation of a pendent phenyl ring and correlates well with efficacy for cytokine inhibition in human whole blood. A hypothetical binding mode is proposed, invoking an expanded ligand binding pocket resembling that of arylpyrazole-bound GR structures. Two compounds of close structural similarity (35 and 37; BMS-776532 and BMS-791826, respectively) have been found to maintain distinct and consistent levels of partial agonist efficacy across several assays, displaying anti-inflammatory activity comparable to that of prednisolone 2 in suppressing cytokine production in whole blood and in rodent models of acute and chronic inflammation.
Subject(s)
Anti-Inflammatory Agents, Non-Steroidal/chemical synthesis , Heterocyclic Compounds, 3-Ring/chemical synthesis , Receptors, Glucocorticoid/agonists , Thiadiazoles/chemical synthesis , Alkaline Phosphatase/biosynthesis , Animals , Anti-Inflammatory Agents, Non-Steroidal/chemistry , Anti-Inflammatory Agents, Non-Steroidal/pharmacology , Arthritis, Experimental/drug therapy , Cell Line, Tumor , Drug Partial Agonism , Edema/drug therapy , Glutamate-Ammonia Ligase/biosynthesis , Heterocyclic Compounds, 3-Ring/chemistry , Heterocyclic Compounds, 3-Ring/pharmacology , Humans , In Vitro Techniques , Interleukin-1beta/blood , Male , Models, Molecular , Rats , Rats, Inbred Lew , Rats, Sprague-Dawley , Receptors, Glucocorticoid/genetics , Receptors, Glucocorticoid/metabolism , Response Elements , Stereoisomerism , Structure-Activity Relationship , Thiadiazoles/chemistry , Thiadiazoles/pharmacology , Transcription Factor AP-1/genetics , Transcription Factor AP-1/metabolism , Transcriptional Activation , Tumor Necrosis Factor-alpha/blood , Tyrosine Transaminase/biosynthesisABSTRACT
A series of 2,2-dimethyl-3,3-diphenyl-propanamides as novel glucocorticoid receptor modulators is reported. SAR exploration led to the identification of 4-hydroxyphenyl propanamide derivatives displaying good agonist activity in GR-mediated transrepression assays and reduced agonist activity in GR-mediated transactivation assays. Compounds 17 and 30 showed anti-inflammatory activity comparable to prednisolone in the rat carrageenan-induced paw edema model, with markedly decreased side effects with regard to increases in blood glucose and expression of hepatic tyrosine aminotransferase. A hypothetical binding mode accounting for the induction of the functional activity by a 4-hydroxyl group is proposed.
Subject(s)
Amides/pharmacology , Receptors, Glucocorticoid/agonists , Amides/chemistry , Animals , Models, Molecular , RatsABSTRACT
The first stereoselective synthesis of the hexahydroimidazo[1,5b]isoquinoline (HHII) scaffold as a surrogate for the steroidal A-B ring system is described. The structure-activity relationships of the analogs derived from this scaffold show that the basic imidazole moiety is tolerated by the glucocorticoid receptor (GR) in terms of binding affinity, although the partial agonist activity in the transrepressive assays depends on the substitution pattern on the B-ring. More importantly, most compounds in the HHII series bearing a tertiary alcohol moiety on the B-ring are either inactive or significantly less active in inducing GR-mediated transactivation, thus displaying a "dissociated" pharmacology in vitro.
Subject(s)
Isoquinolines/chemical synthesis , Isoquinolines/pharmacology , Receptors, Glucocorticoid/agonists , Dexamethasone/pharmacology , E-Selectin/genetics , E-Selectin/metabolism , Genes, Reporter , HeLa Cells , Humans , Isoquinolines/chemistry , Lung/cytology , Lung/drug effects , Molecular Structure , Promoter Regions, Genetic , Structure-Activity Relationship , Transcription, Genetic , Transcriptional ActivationABSTRACT
A series of dihydro-9,10-ethano-anthracene-11-carboxamides as novel glucocorticoid receptor modulators is reported. SAR exploration identified compounds from this series displaying a promising dissociation profile in discriminating between transrepression and transactivation activities. 17a is a partial agonist of GR-mediated transactivation which elicits potent and efficacious transrepression in reporter gene assays. A hypothetical binding mode is provided which accounts for the induction of functional activity by a bridgehead methyl group.
